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Screening of Tropical Fruits for Anti-
Inflammation Activity in Vitro in South
Kalimantan Indonesia
Farizka Erianti1, Alvisha Nadhila R.
1, Adiba
1, and Eko Suhartono
2
1School of Medicine Lambung Mangkurat University, Banjarmasin, South Kalimantan, Indonesia.
2Medical Chemistry/Biochemistry Department, School of Medicine Lambung Mangkurat University, Banjarmasin,
South Kalimantan Indonesia
Email: {fariskaerianti, ekoantioxidant}@gmail.com
Abstract—In recent years many studies revealed that fruits
contain high antioxidant and anti-inflammatory activity. South Kalimantan Indonesia is a tropical country that has a
variety of fruits that are consumed for food and health. The
present study was aimed to evaluate the in vitro anti
inflammation activity of Annona muricata, Averrhoa
carambola and Ananas comosus merr. The anti-
inflammatory activities was determined by inhibition
protein denaturation method. Result of this study revealed
there is inhibitory action on protein denaturation. Averrhoa
carambola extract has shown better inhibition of BSA
denaturation at any concentration compared to Annona
muricata and Ananas comosus merr extract. Our
investigation suggest that the Annona muricata, Averrhoa
carambola, and Ananas comosus merr extracts possess
significant anti-inflammatory activity. Among them
Averrhoa carambola extract was found to be more potent
than the Anona muricata and Ananas comosus merr extract.
Index Terms—ananas comosus merr, annona muricata, anti-
inflammation activity, averrhoa carambola
I. INTRODUCTION
Inflammation is the complex biological response of
vascular tissues to harmful stimuli including pathogens,
irritants, or damaged cells. It is a protective mechanism
by the organism to remove the injurious stimuli as well as
initiate the healing process for the tissue. The process of
inflammation is required in healing of wounds. However,
if inflammation is unchecked, leads to onset of diseases
like rhinnorhoea, rheumatoid arthritis and atherosclerosis.
Acute inflammation is characterized by classical signs
like edema, erythema, pain, heat, and primarily loss of
function [1].
Early inflammatory changes in damaged tissues are
known to release various biologically active materials
from polymorph nuclear leukocytes, lysosomal enzymes
and others. The vascular effects are mediated by
prostaglandins, kinins and vasoactive amines (histamine)
released by mast cells. Acute models are designed to test
drugs that modulate erythema, measurement of local pain,
antipyretic activity, local analgesic action and rat paw
Manuscript received July 10, 2014; revised October 24, 2014.
edema, changes in vascular permeability, leukocyte
migration and chemotaxis, phagocytosis-
polymorphonuclear leucocytes and other phagocytic cells
[2].
All the steroidal and non-steroidal anti-inflammatory
drugs (NSAIDs) although effective, cause undesirable
and side effects. Thus, the development of a safer and
efficacious alternative is needed. Natural products serve
as an important source of therapeutically effective
medicines [1].
In recent years many studies revealed that fruits
contain high antioxidant and anti-inflammatory activity.
[3]. Medicinal plants, which form the backbone of
traditional medicine, have in the last few decades been
the subject of very intense pharmacological studies. Plant
derived drugs serve as a prototype to develop more
effective and less toxic medicines [4]. The curative
properties of medicinal plants are perhaps due to the
presence of various secondary metabolites such as
alkaloids, flavonoids, phenols, saponins, sterols etc [5].
There is a growing attention in correlating the
phytochemicals of a medicinal plant with its
pharmacological activity [6].
South Kalimantan Indonesia is a tropical country that
has a variety of fruits that are consumed for food and
health. For example Sirsak (Annona muricata). Annona
muricata is a member of the family of custard apple tree
called annonaceae and a species of the genus Annona
known mostly for its edible fruits annona [7]. Annona
muricata produces fruits that are usually called sour sop
due to its slightly acidic taste when ripe. The creamy and
delectable flesh of the fruit consist of 80% water, 1%
protein, 18% carbohydrates and fair amount of vitamins
B, B2 and C, potassium and dietary fiber [8], [9]. A.
muricata ethnomedicinal use, especially for inflammation,
rheumatism and neuralgy [9].
Besides Sirsak, there is Belimbing (Averrhoa
carambola) and Nenas (Ananas comosus merr). Averrhoa
carambola L. (Oxalidaceae) is an Asian tree. This tree is
also known as the star fruit tree and is commonly used to
treat headaches, vomiting, coughing and hangovers.
Furthermore, it is used as an appetite stimulant, a diuretic,
and as an antidiarrheal and febrifugal agent [10].
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Journal of Medical and Bioengineering Vol. 4, No. 5, October 2015
©2015 Engineering and Technology Publishingdoi: 10.12720/jomb.4.5.407-411
Sripanidkulchai B. et al (2002) reported that Averrhoa
carambola inhibited carrageenan-induced rat paw
inflammation In another investigation, Cabrini DA. et al
(2010) reported that upon topical application of the
ethanolic extract of Averrhoa carambola leaf and its
butanol, ethyl acetate and hexane fractions, croton oil-
induced ear edema and cellular migration in mice were
both reduced effectively [11].
Ananas comosus (L.) Merrill is belonging to the family
Bromeliaceae and it is an important tropical and
subtropical plant widely cultivated in the tropical areas of
the world. Its fruit is consumed fresh or canned as a
commercial product in many countries. Pineapple has
also been known for a number of beneficial biological
activities such as antioxidative, anti-browning, anti-
inflammatory and anti-platelet activities. The enzyme
complex of A. comosus called bromelain is known for its
clinical applications particularly modulation of tumor
growth, blood coagulation and ant inflammatory effect.
Pineapple has been extensively used in foods or for health
benefits[12].
The present study was aimed to evaluate the in vitro
anti-inflammation activity of Annona muricata, Averrhoa
carambola and Ananas comosus merr.
II. MATERIAL AND METHODS
A. Plant Materials and Preparation of Fruits Extracts
Fruit of Annona muricata, Averrhoa carambola, and
Ananas comosus merr were obtained from Banjarbaru
Traditional Market, South Kalimantan, Indonesia. The
fruit with seed were separated. Fruits were washed with
distilled water. The collected fruits were cut into small
pieces and blended by juicer.
B. In Vitro Anti-Inflammation Activity
Test solution (0,5 ml) consist of 0,45 ml of Bovine
Serum Albumin (BSA) (5% w/v aqueous solution) and
0,05 ml of test solution (250 g/L).
Test control solution (0,5 mL) consists of 0,45 ml of
BSA (5% w/v aqueous solution) and 0,05 ml of distilled
water.
Product control solution (0,5 ml) consists of 0,45 ml of
distilled water and 0,05 ml of test solution (250 g/ml).
Standard solution (0,5 ml) consists of 045 ml of BSA
(5% w/v aqueous solution) and 0,05 ml diclofenac
sodium (250 g/ml).
All the above solution were adjusted to pH 6,3 using 1
N hydrochloric acid. The samples were incubated at 37OC
for 20 min and the temperature was increased to keep the
samples at 57OC for 3 min. After cooling 2,5 ml of
phosphate buffer saline was added to the above solutions.
The absorbance was measured using UV visible
spectrophotometer at 416 nm. The percentage inhibition
of protein denaturation was calculated as, [13]
(1)
The control represents 100% protein denaturation. The
results were compared with diclofenac sodium
(250g/ml).
C. Statistical Analysis
For the anti-inflammation activity, Inhibitory
Concentration 50% (IC50) was measured. The IC50 was
calculated from the linear curve using microsoft excell
2007 and obtained by plotting the percentage of
inhibition versus the concentrations.
III. RESULTS
Anti-inflammation activity of three fruits extracts was
measured by protein denaturation inhibition method. The
results are showed in Fig. 1-Fig. 3 and Table I.
Annona muricata
y = 1.9978x + 16.218
R2 = 0.6947
0
20
40
60
80
0 10 20 30 40
Extract Concentration (%)
Po
ten
ial I
nh
ibit
ion
(%
)
Figure 1.
Anti-Inflammation
Activity of Annona muricata
Averrhoa carambola
y = 2.5331x + 19.866
R2 = 0.7088
0
20
40
60
80
100
120
0 10 20 30 40
Extract Concentration (%)
Po
ten
ial I
nh
ibit
ion
(%
)
Figure 2.
Anti-Inflammation
Activity of Averrhoa carambola
Ananas comosus merr
y = 0.7487x + 1.487
R2 = 0.9636
0
5
10
15
20
25
30
0 10 20 30 40
Extract Concentration (%)
Po
ten
ial I
nh
ibit
ion
(%
)
Figure 3.
Anti-Inflammation Activity of Ananas comosus merr
% Inhibition 100 – 0.D test solution – 0.D of product solutionb
0,D of test control
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Journal of Medical and Bioengineering Vol. 4, No. 5, October 2015
©2015 Engineering and Technology Publishing
The three fruits extracts inhibited the denaturation of
Bovine Serum Albumin (BSA). The degree of inhibition
of BSA denaturation increased with the increasing in the
concentration of extracts. As shown in Fig. 1-Fig. 3 and
Table I among the three extracts under the study
Averrhoa carambola extract (from 71,07% to 82,00%)
has shown better inhibition of BSA denaturation at any
concentration compared to Annona muricata and Ananas
comosus merr extract. The standard drug Diclofenac
sodium showed 85,65% maximum inhibition of
denaturation at 30 μg/ml concentration.
TABLE I. EFFECT OF SELECTED FRUITS ON PROTEIN DENATURATION
Sample Concentration (%) % Inhibition (%) IC 50
Annona muricata
0
10 20
30
0,00
57,63 61,96
65,15
16,19
Averrhoa
carambola
0 10
20 30
0,00 71,07
78,38 82,00
11,81
Ananas comosus merr
0
10 20
30
0,00
11,69 15,49
23,69
64,71
Natrium
Diclofenac
0 10
20 30
0,000 70,16
77,22 85,65
11,87
Result of this study also revealed he concentration
required for 50% inhibition (IC50). IC50 was found to be
16,19 for Annona muricata, 11,81 for Acerrhoa
carambola, and 64,71 for Ananas comosus merr extracts
(Table I). Meanwhile, IC50 for sodium diclofenac is
11,87. The results showed that, the closest IC50 values
IC50 values of diclofenac sodium is Averrhoa carambola
extracs.
IV. DISCUSSION
There are certain problems in using animals in
experimental pharmacological research, such as ethical
issues and the lack of rationale for their use when other
suitable methods are available or could be investigated.
Hence, in the present study the protein denaturation
bioassay was selected for in vitro assessment of
antiinflammatory property of three selected tropical fruits
extract [14].
Denaturation of tissue proteins is one of the well-
documented causes of inflammatory and arthritic diseases.
Production of auto antigens in certain arthritic diseases
may be due to denaturation of proteins in vivo[15].
Agents that can prevent protein denaturation therefore,
would be worthwhile for anti-inflammatory drug
development. The increments in absorbances of test
samples with respect to control indicated stabilization of
protein i.e. inhibition of protein (albumin) denaturation
by extracts and reference drug diclofenac sodium [16].
From the IC50 values it becomes evident that one of the
extracts (averrhoa carambola) was more active than
diclofenac sodium, being effective in lower
concentrations.
Inflammation is natural response of the mammalian
body to a variety of hostile agents including parasites,
pathogenic microorganism, toxic chemical substances
and physical damage to tissue. The process associated
with the inflammatory response are complex but
important aspects which have been exploited for
screening of anti inflammatory compound [17]. Although
steroidal anti-inflammatory drugs and NSAIDs are
currently used to treat acute inflammation, these drugs
have not been entirely successful in curing chronic
inflammatory disorders while such compounds are
accompanied by unexpected side effects. Therefore, there
is an urgent need to find safer anti-inflammatory
compounds [18]. Traditional medicine has used extracts
of different plants for the treatment of a wide variety of
disorders including acute and chronic inflammation [19].
It is commonly accepted that in a situation of oxidative
stress, reactive oxygen species (ROS) such as superoxide,
hydroxyl and peroxyl radicals are generated. The ROS
play an important role in the pathogenesis of various
serious diseases, such as neurodegenerative disorders,
cancer, cardiovascular diseases, atherosclerosis, cataracts,
and inflammation. The mechanism of inflammation
injury is attributed, in part, to release of reactive oxygen
species from activated neutrophils and macrophages [20].
This over production leads to tissue injury by damaging
macromolecules [20]. The main cellular components
susceptible to damage by ROSare lipids (peroxidation of
unsaturated fatty acids in cell membrane), proteins
(denaturation), carbohydrates and nucleic acids [14].
Both inflammation and free radical damage are inter-
related aspects that influence each other. As said above
proteins are susceptible to undergo denaturation by
formation of free radicals and the mechanism of
inflammation injury is attributed, in part, to release of
ROS from activated neutrophil and macrophages Thus
free radicals are important mediators that provoke or
sustain inflammatory processes and consequently, their
neutralization by antioxidants and radical scavengers can
attenuate inflammation [14].
Fruits and vegetables contain significant levels of
biologically active components that impart health benefits
beyond basic nutrition. They are a major source of dietary
antioxidants that increase the plasma antioxidant capacity.
Several classes of antioxidant dietary compounds have
been suggested to present health benefits, and there are
evidences that consumption of these products leadsto a
reduction of the expression of various inflammation
reaction [21].
Annona muricata is one of the tropical fruits that
demonstrate antioxidant properties [22]. Annona
muricata of the family Annonaceae, commonly known as
Sirsak, is a well known medicinal tree with anti-bacterial,
antiviral, molluscicidal, anti-oxidative stress and diuretic
properties [23].
Fruits of the genus Annona contain a considerable
amount of polyphenolic compounds. These compounds
are antioxidants and help to prevent diseases associated
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Journal of Medical and Bioengineering Vol. 4, No. 5, October 2015
©2015 Engineering and Technology Publishing
with oxidative stress, such as cancer, atherosclerosis and
neurodegener ative diseases. A mechanism for
antioxidant activity of natural compounds is the inhibition
and suppression of the formation of reactive species
either by inhibiting enzymes or by chelation of the trace
elements involved in the formation of free radicals.
Antioxidants also participate as free radical scavengers by
regulating or protecting the endogenous antioxidant
defense [24, [25].
Averrhoa carambola is a small, attractive,
multistemmed, slow growing evergreen tree with a short
trunk or a shrub, 5-7m of height or rarely, 10m high,
spreading 20-25 ft in diameter The fruits are green when
small and unripe but turn to yellow or orange when
matured and ripe [11]. Phytochemistry studies have
shown that the fruit of A. carambola is rich in
antioxidants, especially polyphenolic compounds, which
act against reactive oxygen species [26]. Besides
polyphenolic, the fruit of A. Carambola contain saponin,
alkaloid, flavonoid, and tannin. Sripanidkulchai B. et al
(2002) reported that Averrhoa carambola inhibited
carrageenan-induced rat paw inflammation [11].
Pineapple (Ananas comosus Merr) is an important
tropical and subtropical plant widely cultivated in the
tropical areas of the world. Its fruit is consumed fresh or
canned as a commercial product in many countries.
Pineapple has also been known for a number of beneficial
biological activities such as antioxidative, anti-browning,
anti-inflammatory and anti-platelet activities [27].
Pineapple fruit is considered a highly nutritious fruit
because it contains a high level of vitamin C, a natural
antioxidant which may inhibit the development of major
clinical conditions including heart disease and certain
cancers. The fruit also contains phenolic compounds and
β-carotene, which constitute natural sources of
antioxidants [28].
As said above neutralization by antioxidants and
radical scavengers can attenuate inflammation. Thus the
presence of anti-inflammatory activity can be attributed
to the phytochemical compounds in three selected fruit
extracts which acts as an antioxidant or radical scavenger.
Further definitive studies are necessary to ascertain the
mechanism and costituents behind its anti-inflammatory
actions.
V. CONCLUSION
Our investigation clearly demonstrates that the Annona
muricata, Averrhoa carambola, and Ananas comosus
merr extracts possess significant anti-inflammatory
activity. Among them Averrhoa carambola extract was
found to be more potent than the Anona muricata and
Ananas comosus merr extract. Futher studies are
recommended to isolate the active principle responsible
for these activities.
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Fariska Erianti was born in Banjarmasin, South Kalimantan, Indonesia, in August 1993.
She currently study medical doctor in School of Medicine Lambung Mangkurat University,
Banjarbaru, Indonesia.
Alvisha Nadilla R. was born in Banjarmasin,
South Kalimantan, Indonesia, in February 1995. She currently study medical doctor in
School of Medicine Lambung Mangkurat University, Banjarbaru, Indonesia.
Adiba was born in Banjarmasin, South
Kalimantan, Indonesia, in March 1994. She currently study medical doctor in School of
Medicine Lambung Mangkurat University,
Banjarbaru, Indonesia.
Eko Suhartono was born in Surabaya,
Indonesia, in September 1968. He received his
Drs. and M.Sc degree in 1991 and 1998 from
Surabaya Technic Institute, Surabaya, East Java and Gadjah Mada University, Yogyakarta,
Indonesia. He currently study environmental science and technology graduate program in
Brawijaya University, Malang, Indonesia. His
research is mainly focused on free radical and natural product antioxidant, ecotoxicology.
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Journal of Medical and Bioengineering Vol. 4, No. 5, October 2015
©2015 Engineering and Technology Publishing
